Reexamination of the polymerization of pyridoxylated hemoglobin with glutaraldehyde

Biopolymers
M A MariniA Zegna

Abstract

Pyridoxylated adult human hemoglobin (HbAo) was prepared using a one molar equivalent of pyridoxal 5-phosphate (PLP) per heme and reduced with either NaCNBH3 or NaBH4. A separate sample was pyridoxylated and passed through a mixed-bed ion exchange column without reduction. All three preparations had a P50 of 29 +/- 2 torr and a cooperativity of n = 2.4 +/- 0.1. These preparations, in both the oxy and deoxy forms, were then treated with 7 equivalents of glutaraldehyde per tetramer at pH 6.8 at 4 degrees C and at room temperature. The polymerization invariably reduced the P50 to 18 +/- 2 torr with Hill coefficients of less than 2. These solutions, with or without further reduction using NaCNBH3, all retained the PLP in differing amounts (2-3 moles/tetramer). Methemoglobin concentrations were increased during the polymerization reaction. The normal pyridoxylation procedure, using sodium borohydride reduction, resulted in a number of different molecular species. Polymerization with glutaraldehyde caused a further proliferation of molecular species that could not be separated by anion exchange chromatography or by isoelectric focusing. The extent of polymerization, estimated by gel exclusion chromatography and SDS polyacrylamide gel...Continue Reading

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Citations

May 1, 1992·Biopolymers·D KlettE Dellacherie
Jan 3, 2007·Seminars in Hematology·Robert M Winslow
Mar 12, 2008·Antioxidants & Redox Signaling·Caroline Gaucher, Patrick Menu
Jan 1, 1995·Vox Sanguinis·J E Ogden, S L Mac Donald
Aug 24, 2004·Artificial Organs·Robert M Winslow
Sep 1, 1996·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·G MarchandK Burhop
Jun 17, 2008·Biochimica Et Biophysica Acta·Robert M Winslow
Aug 1, 1996·The Journal of Laboratory and Clinical Medicine·R M Winslow
Jun 8, 2002·Biochemical and Biophysical Research Communications·Tao Hu, Zhiguo Su
Jul 28, 2004·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·Wen-Hsiang ChangHsing-Wen Sung
Jan 1, 1995·Annales Françaises D'anesthèsie Et De Rèanimation·J G Riess

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